Unrolling worms: Genome Editing to Rewrite Roller Phenotypes in C. elegans

The gain-of-function allele rol-6 ( su1006 ) has been used as a co-injection marker to visually identify transgenic progeny following microinjection in Caenorhabditis elegans . The rol-6 (gf) allele yields a clear, visual roller phenotype; however, affected worms are twisted along the anterior-posterior axis, obscuring tissues and complicating visual analyses. We deployed CRISPR/Cas9 to "unroll" transgenic worm strains harboring rol-6 (gf) . We discovered that our successfully unrolled strains introduced additional nucleotides into the rol-6 (gf) loci, rendering the rol-6 (gf) copies inactive. Our results indicate that genome engineering can be easily deployed to modify existing transgenic worm strains and could be applied to other gain-of-function co-injection markers.